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The insulin-degrading enzyme is an allosteric modulator of the 20S proteasome and a potential competitor of the 19S

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Abstract

The interaction of insulin-degrading enzyme (IDE) with the main intracellular proteasome assemblies (i.e, 30S, 26S and 20S) was analyzed by enzymatic activity, mass spectrometry and native gel electrophoresis. IDE was mainly detected in association with assemblies with at least one free 20S end and biochemical investigations suggest that IDE competes with the 19S in vitro. IDE directly binds the 20S and affects its proteolytic activities in a bimodal fashion, very similar in human and yeast 20S, inhibiting at (IDE) ≤ 30 nM and activating at (IDE) ≥ 30 nM. Only an activating effect is observed in a yeast mutant locked in the “open” conformation (i.e., the α-3ΔN 20S), envisaging a possible role of IDE as modulator of the 20S “open”–”closed” allosteric equilibrium. Protein–protein docking in silico proposes that the interaction between IDE and the 20S could involve the C-term helix of the 20S α-3 subunit which regulates the gate opening of the 20S.

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Acknowledgements

The authors thank Prof. Michael Glickman and Prof. Peter van Endert, for several fruitful discussions. We are grateful to Prof. Michael Groll for the generous gift of wt y20S and α-3ΔN mutant proteasome. The financial support from the Italian Ministry of University and Research (MiUR PRIN20157WZM8 to G.G. and S.M.) is gratefully acknowledged.

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  1. Diego Sbardella and Grazia R. Tundo contributed equally to this paper.

Authors and Affiliations

  1. Department of Clinical Sciences and Translational Medicine, University of Roma Tor Vergata, Via Montpellier 1, 00133, Rome, Italy

    Diego Sbardella, Grazia R. Tundo, Chiara Ciaccio, Paola Cozza, Stefano Marini & Massimo Coletta

  2. Interuniversitary Center for the Research on the Chemistry of Metals in Biological Systems, Bari, Italy

    Diego Sbardella, Grazia R. Tundo, Chiara Ciaccio, Stefano Marini & Massimo Coletta

  3. Interdepartmental Center for TeleInfrastructures, University of Roma Tor Vergata, Rome, Italy

    Diego Sbardella, Paola Cozza, Stefano Marini & Massimo Coletta

  4. Department of Chemistry, Aarhus University, Aarhus, Denmark

    Andrea Coletta

  5. Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, Toulouse, France

    Julien Marcoux, Efthymia Ioanna Koufogeorgou & Marie-Pierre Bousquet-Dubouch

  6. Institute of Biostructures and Bioimaging, National Research Council, Catania, Italy

    Anna M. Santoro & Danilo Milardi

  7. Department of Chemistry, University of Catania, Catania, Italy

    Giuseppe Grasso

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  1. Diego Sbardella
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Sbardella, D., Tundo, G.R., Coletta, A. et al. The insulin-degrading enzyme is an allosteric modulator of the 20S proteasome and a potential competitor of the 19S. Cell. Mol. Life Sci. 75, 3441–3456 (2018). https://doi.org/10.1007/s00018-018-2807-y

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